Mycobacterium tuberculosis and other Mycobacterium species are major pathogens around the world. They are part of a larger group of so-called fastidious pathogens that are difficult to study because, for a wide range of reasons, they are difficult to culture. Phenotype MicroArrayTM (PM) technology is a tool that can aid in understanding the physiological and metabolic properties of fastidious pathogens. It enables a scientist to scan nearly 2,000 phenotypes of a microbial cell line in a single experiment. This technology can now be used with most aerobic gram negative/positive species of interest in human health and more than 70 scientific publications and presentations demonstrate how it can be applied in a wide range of scientific investigations. However, a number of fastidious genera are currently not amenable to PM analysis. One principal goal for this STTR project is to adapt PM technology for these fastidious aerobic, microaerophilic, and anaerobic bacterial species, with special emphasis on those important in human disease and/or biodefense considerations. The genera we will focus on include agents of lung, cutaneous, and tissue infections (Mycobacterium, Nocardia, Legionella), microaerophilic gastro-intestinal pathogens (Helicobacter, Campylobacter, Arcobacter, Wolinella), and important colonizers of the colon and vagina (Bacteroides, Clostridium, Lactobacillus, Escherichia). These were selected as highest priority because we have received requests from more than 40 scientists seeking to utilize PM technology in their studies of these microorganisms. This level of scientific interest documents the need and potential for commercialization of the technology that we are proposing to develop. A second goal is to expand the capabilities of the Biolog Microbial Identification System, which is now installed in about half of US State Public Health Labs and is frequently used to identify less common or unusual microbial pathogens. Most of these genera are also not currently included in Biolog's Identification Database. However, with success in development efforts proposed here, the Database will be expanded to include these important fastidious species. As a third goal, our collaborator at Texas A&M University will focus on the most important genus, Mycobacterium, and use knockout strains with PM technology to study the function of genes that are unique to the metabolism and drug resistance of this genus. If successful, this aspect of the project will aid efforts toward the development of new or more effective anti-mycobacterial drug therapies. This project will directly benefit public health by providing a technology for efficient simultaneous testing of hundreds to thousands of cellular phenotypes. It will be applied in hospital diagnostic work, environmental diagnostic work, bioterrorism monitoring, new drug development, toxicology studies, and in many areas of basic biology research.